Transverse Microradiography Research Articles

In vitro near-infrared imaging of natural secondary caries.

Secondary caries stands as the leading reason for the failure of composite restorations and dentists spend more time replacing existing restorations than placing new ones. Current clinical strategies, and even modern visible light methods designed to detect decay, lack the sensitivity to distinguish incipient lesions, are confounded by staining on the surface and within the tooth, or are limited to detecting decay on the tooth surface. Near-IR (NIR) imaging methods, such as NIR reflectance and transillumination imaging, and optical coherence tomography are promising strategies for imaging secondary caries. Wavelengths longer than 1300-nm avoid interference from stain and exploit the greater transparency of sound enamel and dental composites, to provide increased contrast with demineralized tissues and improved imaging depth. The purpose of this study was to determine whether NIR transillumination (λ=1300-nm) and NIR cross-polarized reflectance (λ=1500-1700-nm) images can serve as reliable indicators of demineralization surrounding composite restorations. Twelve composite margins (n=12) consisting of class I, II & V restorations were chosen from ten extracted teeth. The samples were imaged in vitro using NIR transillumination and reflectance, polarization sensitive optical coherence tomography (PS-OCT) and a high-magnification digital visible light microscope. Samples were serially sectioned into 200-μm slices for histological analysis using polarized light microscopy (PLM) and transverse microradiography (TMR). The results presented demonstrate the utility of NIR light for detecting recurrent decay and suggest that NIR images could be a reliable screening tool used in conjunction with PS-OCT for the detection and diagnosis of secondary caries.

Evaluation of caries progression in dentin treated by fluoride-containing materials using an in-air micro-PIGE and micro-PIXE measurement system

It is well-known that fluorine (F) is involved in the progression of caries. The evaluation of caries progression has conventionally been based on the change in mineral content using transverse microradiography (TMR). The purpose of this study was to evaluate the progression of dentinal caries by the change in calcium (Ca) content using Particle-Induced Gamma-ray Emission/Particle-Induced X-ray Emission (PIGE/PIXE) techniques at the Wakasa Wan Energy Research Center. We also assessed the relationship between caries progression rate and the concentration of F penetration into dentin from dental fluoride-containing materials (FCMs). Dentin sections of six extracted human teeth were prepared to obtain various amounts of F uptake using three types of FCMs. F and Ca distribution of specimens were obtained using PIGE/PIXE techniques. After evaluation, the specimens were immersed in 10ml of demineralizing solution (pH 4.5) to simulate caries attack. To estimate caries progression rates, the same portions of the specimens were evaluated after caries attack treatment using PIGE/PIXE. A negative correlation between the F uptake in dentin and the rate of caries progression was observed. Therefore, caries progression in dentin was reduced by increasing the amount of F uptake from FCMs. This demonstrates that PIGE/PIXE techniques are valuable for estimating caries progression rates.

Formation and characterization of hypermineralized zone beneath dentine lesion body induced by topical fluoride in-vitro

ObjectiveThis in-vitro study aimed to evaluate and characterize the hypermineralized zone (Hyper-zone) formed beneath the remineralized dentine lesion body by transverse microradiography (TMR), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDS). DesignDemineralized bovine dentine specimens were treated with fluoride solutions (APF, NaF) and remineralized for 2–4 weeks. Then thin sections were prepared to characterize the Hyper-zone by TMR, EDS. Fractured specimen surfaces were observed by SEM. ResultsTMR analysis revealed a higher mineral density at Hyper-zone than that of sound dentine (48vol%) ranging from 50 up to 61vol% and the thickness ranging from 197 to 344μm for 4-week specimens, while specimens without fluoride treatment did not show Hyper-zone. SEM pictures at Hyper-zone showed no evident crystal-like deposits in dentinal tubules and no notable difference when compared to that in sound dentine. EDS analysis demonstrated higher concentrations of Ca and P at Hyper-zone than those in sound dentine, which corresponded to the TMR profile, while the magnesium (Mg) concentration was low at this zone. ConclusionsDemineralized dentine lesions exposed to fluoride and remineralization treatments exhibited Hyper-zone beneath the lesion body, in which the mineral density was higher than that of sound dentine. Possible mechanism for the formation of Hyper-zone was discussed by assuming removal of mineral regulators such as Mg and other organic substances from sound dentine during de-/remineralization processes.

Biofilm Layers Affect the Treatment Outcomes of NaF and Nano-hydroxyapatite

During caries formation, dental biofilms function not only as acid producers but also as reservoirs and diffusion barriers for active caries-preventive components. The aim of this study was to investigate the influence of biofilms as a stagnant layer on the efficacy of NaF and nano-hydroxyapatite (nHA). Biofilms of Streptococcus mutans C180-2 were formed on the surfaces of artificially demineralized enamel in an active attachment biofilm model. After 2 days of biofilm formation, the model was subjected to a pH-cycling schedule, together with a control group without biofilms. Specimens were treated for 5 min twice daily with water, a 10% nHA slurry, or 18.4 mM NaF. At the end of the pH-cycling period, the biofilms were removed for the determination of the viable counts, the lactic acid production, and the calcium content. The mineral changes in the demineralized enamel blocks were analyzed by transversal microradiography. No differences in the biofilm viable counts and lactic acid production were found in the different treatment groups. The mean calcium content of the biofilms in the nHA group was 60.7 ± 15.3 mmol/g wet weight, which was approximately 8-fold higher than in the other 2 groups. The application of NaF resulted in net remineralization, but in the presence of a biofilm, net demineralization was observed. In contrast, nHA treatment reduced further demineralization compared with the water treatment, but the presence of a biofilm enhanced this effect. In conclusion, the presence of biofilms clearly influenced the treatment outcomes of anticaries products. Biofilms could either enhance or impede their efficacy. This result implies that biofilms should be included in the in vitro tests for the preclinical screening of caries-protective agents.

Control of white spot lesion adjacent to orthodontic bracket with use of fluoride varnish or chlorhexidine gel.

The aims of this study were to compare the effectiveness of fluoride varnish and chlorhexidine gel in controlling white spot lesions (WSLs) adjacent to orthodontic brackets and to compare the ability of Quantitative Light-Induced Fluorescence (QLF) to measure mineral uptake with that of transverse microradiography (TMR). Thirty premolars with artificially induced WSLs were randomly assigned to three groups: (1) two applications of 5% NaF-varnish (F), with one-week interval, (2) two applications of 2% chlorhexidine gel (CHX), with one-week interval, and (3) control (CO), no treatment. QLF was used to measure changes in fluorescence before and after caries induction, 1 week after each application and 1, 2, and 3 months after the last application of F or CHX. TMR was performed to quantify lesion depth and mineral content after caries induction to evaluate the effects of F, CHX, and CO 3 months after the last application of agents. The data were analyzed by repeated measures ANOVA and Tukey's test. All treatments increased the mineral content during the experimental period; however, F induced faster remineralization than CHX. The correlation between QLF and TMR was significantly moderate. Two applications of fluoride varnish or 2% chlorhexidine gel at one-week intervals were effective in controlling WSLs.

Clinical assessment of non carious cervical lesion using swept-source optical coherence tomography.

Non-carious cervical lesions (NCCLs) involve various forms of tooth loss with different etiologies. This study aimed to utilize swept-source optical coherence tomography (SS-OCT) at 1300nm wavelength range in vitro and in vivo to evaluate and clarify the mechanism of NCCLs. In the in vitro phase, a dentin attenuation coefficient (μt ) derived from the SS-OCT signal at NCCL was compared with mineral loss obtained from transverse microradiography (TMR) to determine a μt threshold to discriminate demineralization of cervical dentin in vivo. In the clinical study, 242 buccal surfaces were investigated in 35 subjects. Presence and dimensions of NCCLs, cervical cracking and the degree of demineralization at the exposed cervical dentin were determined using SS-OCT. Dentin demineralization was observed in 69% of NCCLs. SS-OCT results confirm that dentin mineral loss and occlusal attrition were associated with larger NCCLs, and can be considered as an etiological factor in formation and progress of these lesions. (A) We determined the attenuation coeffcient (μt ) threshold of SS-OCT signal for the detection of demineralization (1.21) from in vitro study. DEM: demineralized dentin, sound: sound dentin. (B) Using the μt threshold, we observed NCCLs in vivo to detect the demineralization in cervical dentin. SS-OCT scanning was performed along the red line. (C) SS-OCT image obtained along the red line in B. In SS-OCT, brightness of dentin beneath the NCCL was increased (arrow) compared with intact zone. The cervical dentin was slightly demineralized (μt : 1.25). e: enamel, d: dentin, g: gingiva.

Novel tea polyphenol-modified calcium phosphate nanoparticle and its remineralization potential.

Tea polyphenols (TP) are not only potent antimicrobial and antioxidant agents but also effective modifiers in the formation of nanosized crystals. Since nano-hydroxyapatite (n-HA) is known to enhance remineralization of dental hard tissue, our aims were to synthesize nanosized calcium phosphate particles incorporating TP and to test their potential as caries preventive agent. An ammonia water diffusion method was used to synthesize nanosized calcium phosphate particles (TP-CaP) in the presence of various amounts of TP. The resultant products were characterized by scanning electron microscopy (SEM) and X-ray diffraction (XRD). The remineralization potential of the nano TP-CaP was then investigated in a 12-day pH-cycling model. Nano TP-CaP slurries, at pH 7.0 and pH 5.5, were applied onto preformed enamel lesions 4 × 3 min per day. n-HA slurries at pH 7.0 and pH 5.5 were used as positive controls, and deionized water was served as a negative control. SEM showed nanosized particles were only formed at 27 mg/mL of TP. Further characterization of the nanosized particles revealed the components were amorphous calcium phosphate, HA, and TP. Both surface microhardness and transverse microradiography analyses showed that nano TP-CaP at pH 5.5, but not at pH 7.0, significantly enhanced remineralization, to the same extent as the n-HA controls. Furthermore, significantly higher amount of TP was found in the supernatant of TP-CaP at pH 5.5 than those at pH 7.0. Since TP can inhibit bacterial growth and enzyme activities, the novel nanosized TP-CaP particle, at low pH, is a potential dual-functional-remineralization and antibacteria-product.

Remineralizing Efficacy of a CPP-ACP Cream on Enamel Caries Lesions in situ

The aim of this double-blind, randomized, cross-over in situ study was to compare the remineralizing effects induced by the application of casein phosphopeptide-stabilized amorphous calcium phosphate complexes (CPP-ACP)-containing cream (without fluoride) after the use of fluoride toothpaste with the prolonged use of fluoride toothpaste on enamel caries lesions in situ. During each of three experimental legs of 4 weeks, 13 participants wore intra-oral mandibular appliances with 8 pre-demineralized bovine enamel specimens in the vestibular flanges mimicking either ‘easily cleanable' or ‘proximal' surfaces (n = 312). The three randomly allocated treatments were as follows: (1) application of CPP-ACP-containing cream (GC Tooth Mouse, non-fluoride) after the use of fluoride toothpaste (1,400 ppm NaF; TM), (2) prolonged application of fluoride toothpaste (1,400 ppm NaF; positive control, PC) and (3) prolonged application of fluoride-free toothpaste (negative control, NC). Additionally, one of each of the two flanges was brushed twice daily with the respective toothpaste. The differences in integrated mineral loss as assessed by transversal microradiography were calculated between values before and after the in situ period. Changes in mineral loss were analysed for those pairs of subgroups differing in only one of the three factors (intervention, brushing and position). The PC treatment induced a significantly higher mineral gain compared with the TM and NC treatments. No significant differences between TM and NC for both positions were observed. In conclusion, the additional use of a CPP-ACP-containing cream seems to be less efficacious in remineralizing caries lesions than the prolonged application of fluoride toothpaste. © 2014 S. Karger AG, Basel

Restoration Materials and Secondary Caries Using an In Vitro Biofilm Model

This in vitro study investigated whether restoration materials and adhesives influence secondary caries formation in gaps using a short-term in vitro biofilm model. Sixty enamel–dentin blocks were restored with 6 different restoration materials with or without adhesives (n = 10 per group) with a gap: 1) Clearfil AP-X composite, 2) Clearfil AP-X composite + SE Bond, 3) Clearfil AP-X composite + ProtectBond, 4) Filtek Silorane composite, 5) Filtek Silorane composite + Silorane System adhesive, or 6) Tytin amalgam. Specimens were subjected to an intermittent 1% sucrose biofilm model for 20 days to create artificial caries lesions. Lesion progression in the enamel–dentin next to the different materials was measured in lesion depth (LD) and mineral loss (ML) using transversal wavelength independent microradiography (T-WIM). A regression analysis was used to compare the LD and ML of the different restoration materials at 4 measurement locations: 1 location at the surface of the enamel, 1 location at the wall of the enamel, and 2 locations at the wall of the dentin. A statistically significant effect of AP-X composite with Protect Bond was found for LD and ML at the WallDentin1 location, leading to less advanced wall lesions. An additional finding was that gap size was also statistically significant at the 2 wall locations in dentin, leading to increasing lesion progression with wider gaps. In conclusion, adhesives can influence wall lesion development in gaps. Protect Bond showed significantly less caries progression compared to bare restoration materials or other adhesives in this short-term in vitro biofilm model.

Effect of xylitol varnishes on remineralization of artificial enamel caries lesions in vitro

ObjectivesAnalyse the effect of varnishes containing xylitol alone or combined with fluoride on the remineralization of artificial enamel caries lesions in vitro. MethodsBovine enamel specimens were randomly allocated to 7 groups (n=15/group). Artificial caries lesions were produced by immersion in 30mL of lactic acid buffer containing 3mM CaCl2·2H2O, 3mM KH2PO4, 6μM tetraetil metil diphosphanate (pH 5.0) for 6 days. The enamel blocks were treated with the following varnishes: 10% xylitol; 20% xylitol; 10% xylitol plus F (5% NaF); 20% xylitol plus F (5% NaF); Duofluorid™ (6% NaF, 2.71% F+6% CaF2), Duraphat™ (5% NaF, positive control) and placebo (no-F/xylitol, negative control). The varnishes were applied in a thin layer and removed after 6h. The blocks were subjected to pH-cycles (demineralization—2h/remineralization—22h during 8 days) and enamel alterations were quantified by surface hardness and transversal microradiography. The percentage of surface hardness recovery (%SHR), the integrated mineral loss and lesion depth were statistically analysed by ANOVA/Tukey's test or Kruskal–Wallis/Dunn's test (p<0.05). ResultsEnamel surface remineralization was significantly increased by Duraphat™, 10% xylitol plus F and 20% xylitol plus F formulations, while significant subsurface mineral remineralization could be seen only for enamel treated with Duraphat™, Duofluorid™ and 20% xylitol formulations. Conclusions20% xylitol varnishes seem to be promising alternatives to increase remineralization of artificial caries lesions. Clinical significance: effective vehicles are desirable for caries control. Xylitol varnishes seem to be promising alternatives to increase enamel remineralization in vitro, which should be confirmed by in situ and clinical studies.

Marginal integrity and secondary caries of selectively excavated teeth in vitro

ObjectivesSelective caries removal involves sealing of carious dentine beneath restorations, which might decrease their marginal integrity and increase the susceptibility for secondary caries and microleakage. The present study compared these marginal characteristics of restorations in selectively and completely excavated teeth. MethodsIn 32 premolars, shallow and deep artificial lesions were created on pulpo-axial walls of mesial-distal-occlusal cavities, with mesial and distal margins located in enamel and dentine, respectively. Demineralised dentine was either removed or left before adhesively restoring the teeth (n=8), which were then submitted to thermo-mechanical cycling. The integrity of gingivo-cervical margins was assessed using scanning electron microscopy. In half of each margin, caries was induced adjacent to restorations using a continuous-culture biofilm model, and resulting lesions were evaluated using transversal microradiography. The other half of each margin was used to assess microleakage. ResultsIntegrity or microleakage of margins located in enamel did not differ significantly between groups, and bacterial biofilms did not induce distinct caries lesions in enamel. Dentinal margins in teeth with deep compared with shallow lesions showed a significantly higher proportion of marginal imperfections, gaps and microleakage (p≤0.05, Mann–Whitney/χ2-test). In contrast, neither marginal integrity nor microleakage differed significantly between completely and selectively excavated teeth (p>0.05). Dentinal mineral loss adjacent to restorations did not differ significantly between groups (p>0.80). ConclusionsThe marginal characteristics of restorations were affected by the depth of sealed or excavated lesions, but not by the performed caries excavation. This study did not find selective excavation detrimental for restoration integrity in vitro. Clinical significanceSelective excavation of deep lesions was shown to reduce pulpal risks, whilst leaving caries beneath restorations is feared to compromise the marginal characteristics of the subsequently placed restoration. Based on the present in vitro study, such assumptions cannot be supported.

Saliva substitutes in combination with high-fluoride gel on dentin remineralization.

This study aimed to determine whether the application of a high-fluoride gel could increase the remineralization of subsurface dentin lesions stored in saliva substitutes. Demineralized bovine dentin specimens were stored in mineral water (W), Glandosane (G), or modified Saliva natura (SN). Different treatments were applied twice daily: no treatment, Elmex sensitive mouth rinse (E), ProSchmelz gel (P), Duraphat toothpaste (D), ED, PD, and EPD. Differences in mineral loss were evaluated by transversal microradiography after 2 and 5 weeks. The treatments with E, D, and ED inhibited the mineral loss induced by G and enabled some mineral gain. ProSchmelz was not able to inhibit the demineralizing effect of G. This high-fluoride gel induced an erosive mineral loss in combination with G. The use of ProSchmelz in combination or not with other fluoride products did not increase remineralization of specimens stored in SN or W (p > 0.05). ProSchmelz resulted in an erosion of the specimens stored in W and revealed a lower mineralized surface layer of specimens stored in SN. Topical application of high-fluoride gel reduced the mineral loss induced by G but resulted in an erosion of specimens' surface. In addition, ProSchmelz did not demonstrate beneficial effects in combination with SN on subsurface dentin lesion remineralization. Within the limitations of an in vitro study, it was concluded that the application of a high-fluoride gel did not promote additional effects on remineralization of subsurface dentin lesions in combination with saliva substitutes when compared to products with lower fluoride concentration.

Gap Size and Wall Lesion Development Next to Composite

This in situ study investigated whether there is a relationship between gap size and wall lesion development in dentin next to 2 composite materials, and whether a clinically relevant threshold for the gap size could be established. For 21 days, 14 volunteers wore a modified occlusal splint containing human dentin samples with 5 different interfaces: 4 gaps of 50 µm, 100 µm, 200 µm, or 400 µm and 1 non-bonded interface without a gap. Eight times a day, the splint with samples was dipped in a 20% sucrose solution for 10 minutes. Before and after caries development, specimens were imaged with transversal wavelength-independent microradiography (T-WIM), and lesion depth (LD) and mineral loss (ML) were calculated at the 5 different interfaces. After correction for the confounder location (more mesial or distal), a paired t test clustered within volunteers was performed for comparison of gap widths. Results showed no trend for a relationship between the corrected lesion depth and the gap size. None of the differences in lesion depth for the different gap sizes was statistically significant. Also, the composite material (AP-X or Filtek Supreme) gave no statistically significant differences in lesion depth and mineral loss. A minimum gap size could not be established, although, in a non-bonded interface without a measurable gap, wall lesion development was never observed.

Comparison of Knoop and Vickers surface microhardness and transverse microradiography for the study of early caries lesion formation in human and bovine enamel

ObjectiveThe aims of the present laboratory study were twofold: a) to investigate the suitability of Knoop and Vickers surface microhardness (SMH) in comparison to transverse microradiography (TMR) to investigate early enamel caries lesion formation; b) to compare the kinetics of caries lesion initiation and progression between human and bovine enamel. DesignSpecimens (90×bovine and 90×human enamel) were divided into six groups (demineralization times of 8/16/24/32/40/48h) of 15 per enamel type and demineralized using a partially saturated lactic acid solution. SMH was measured before and after demineralization and changes in indentation length (ΔIL) calculated. Lesions were characterized using TMR. Data were analyzed (two-way ANOVA) and Pearson correlation coefficients calculated. ResultsΔIL increased with increasing demineralization times but plateaued after 40h, whereas lesion depth (L) and integrated mineral loss (ΔZ) increased almost linearly throughout. No differences between Knoop and Vickers SMH in their ability to measure enamel demineralization were observed as both correlated strongly. Overall, ΔIL correlated strongly with ΔZ and L but only moderately with the degree of surface zone mineralization, whereas ΔZ and L correlated strongly. Bovine demineralized faster than human enamel (all techniques). ConclusionsLesions in bovine formed faster than in human enamel, although the resulting lesions were almost indistinguishable in their mineral distribution characteristics. Early caries lesion demineralization can be sufficiently studied by SMH, but its limitations on the assessment of the mineral status of more demineralized lesions must be considered. Ideally, complementary techniques to assess changes in both physical and chemical lesion characteristics would be employed.

Protective benefits of a stabilised stannous-containing fluoride dentifrice against erosive acid damage

To assess the potential of a stabilised stannous (Sn)-containing NaF dentifrice (Oral B/blend-a-Med(®) Pro-Expert), in addition to a number of other marketed European dentifrices formulated with various fluoride actives and two control dentifrices, to protect enamel against erosive acid damage. Cores of human enamel (four per group) were soaked in pooled human saliva, and then treated with a 1:3 slurry (dentifrice:saliva) using a standardised in vitro erosion model (5-day cycling) that includes 10-minute challenges with 1% citric acid applied 60 minutes after each dentifrice treatment. Enamel surface loss was measured using transverse microradiography (TMR). Specimens treated with the Sn-containing NaF dentifrice showed 6.5 μm of surface loss ± 1.2 (SEM), which was not significantly different (P < 0.05, Fisher LSD) from that of a clinically proven, stabilised SnF2 positive control [Crest(®) Pro-Health, 1,100 ppm F as SnF2 : 3.0 μm of surface loss ± 1.1 (SEM)]. The Sn-containing NaF dentifrice and the clinically proven positive control both provided significantly greater protection (P < 0.05, Fisher LSD) compared with all of the other products tested. Enamel loss (SEM) values for other European products and the reference control (active agents) were: Meridol(®) : (1,400 ppm F as AmF + SnF2 ) 12.0 μm (1.47); Colgate(®) Cavity Protection: (1,450 ppm F as SMFP + NaF) 12.9 μm (1.66); Odol med 3(®) (1,400 ppm F as NaF) 14.2 μm (1.49); Elmex(®) (1,400 ppm F as AmF) 14.5 μm (1.76); Colgate(®) Enamel Protect: (1,450 ppm F as NaF + KNO3 ) 16.3 μm (2.02); Lacalut(®) aktiv: (1,400 ppm F as AlF3 ) 18.5 μm (1.71); Sensodyne(®) ProNamel(™) : (1,450 ppm F as NaF + KNO3 ) 20.5 μm (1.26); Crest Cavity Protection (1,100 ppm F as NaF, reference control) 22.00 μm (2.04); and Mentadent(®) : (1,450 ppm F as NaF + Zn citrate) 22.3 μm (0.63). These results support the potential for the stabilised, Sn-containing NaF dentifrice to provide erosion protection benefits that are not significantly different from the positive control benchmark for erosion protection (stabilised SnF2 ), and are significantly better than a broad range of dentifrice formulations available on the European market.

8DSS-Promoted Remineralization of Initial Enamel Caries In Vitro

Peptides containing 8 repeats of aspartate-serine-serine (8DSS) have been shown to promote the nucleation of calcium phosphate from solution into human enamel. Here we tested the ability of 8DSS to promote the remineralization of demineralized enamel in an in vitro model of artificial early enamel caries. Initial caries lesions were created in bovine enamel blocks, which were then subjected to 12 d of pH cycling in the presence of 25 µM 8DSS, 1 g/L NaF (positive control) or buffer alone (negative control). Absorption of 8DSS was verified by X-ray photoelectron spectroscopy. Mineral loss, lesion depth, and mineral content at the surface layer and at different depths of the lesion body were analyzed before and after pH cycling by polarized light microscopy and transverse microradiography. Mineral loss after pH cycling was significantly lower in the 8DSS samples than in the buffer-only samples, and lesions in the 8DSS samples were significantly less deep. Samples treated with 8DSS showed significantly higher mineral content than buffer-only samples in the region extending from the surface layer (30 µm) to the average lesion depth (110 µm). No significant differences were found between the samples treated with 8DSS and those treated with NaF. These findings suggest that 8DSS has the potential to promote remineralization of demineralized enamel.

Chemical alteration by tooth bleaching of human salivary proteins that infiltrated subsurface enamel lesions--experimental study with bovine lesion model systems.

Salivary macromolecules infiltrate white and brown spot enamel lesions and adsorb onto hydroxyapatite. Calcium-binding salivary proteins such as statherin hinder remineralization of these lesions. We assessed whether bleaching agents can remove salivary components that have infiltrated and bound to experimental subsurface lesions in bovine enamel prepared by immersing specimens in acid and then human saliva. Transversal microradiography showed that such demineralized lesions mimicked incipient carious lesions. Bound proteins to the experimental and untreated control specimens were eluted in a stepwise manner with phosphatebuffered saline, 0.4 M phosphate buffer, and 1 M HCl. SDS-PAGE of dialyzed extracts showed that specific salivary proteins bound to the lesions, while virtually no protein bands were detected if the specimens were bleached. Western blotting showed that even statherin, which was more firmly bound than other proteins, was removed. In-office bleaching agent may be useful in treating enamel lesions for removing proteins bound to these lesions.

In vitro progression of artificial white spot lesions sealed with an infiltrant resin.

This study assessed the ability of an infiltrant resin (Icon, DMG Chemisch-Pharmazeutische Fabrik GmbH, Hamburg, Germany) to prevent artificial lesion progression in vitro when used to impregnate white spot lesions and also assessed the effect of saliva contamination on resin infiltration. Enamel specimens (n=252) were prepared and covered with nail varnish, leaving a window of sound enamel. After demineralization (pH 5.0; four weeks), specimens were divided into six groups (n=42 per group): group 1, 2% fluoride gel (positive control); group 2, resin infiltrant; group 3, resin infiltrant + fluoride gel; group 4, no treatment (negative control); group 5, resin infiltrant application after saliva contamination; and group 6, resin infiltrant + fluoride gel after saliva contamination. Specimens from each group were cut perpendicular to the surface, and one-half of each specimen was exposed to a demineralizing solution for another four weeks. The other half was set aside as a record of initial lesion depth and was used later in the determination of lesion progression. Lesion progression and infiltrant penetration were measured using confocal laser scanning microscopy (CLSM) and transverse microradiography (TMR). For lesion depth, based on CLSM, groups 2 and 3 showed the least changes when submitted to demineralization challenge, followed by group 1, then groups 5 and 6, and finally group 4. There were no significant differences between groups 2 and 3 or groups 5 and 6 in their ability to inhibit further lesion progression (p<0.05). Based on TMR, groups 2 and 3 also showed the fewest changes when submitted to demineralization challenge, followed by group 5, then groups 1 and 6, and finally group 4. In terms of mineral loss as measured by TMR, all groups that contained fluoride (groups 1, 3, and 6) show less percentage change in mineral loss than the groups that did not contain fluoride (groups 2, 4, and 5). It can be concluded that infiltrant penetration into early enamel lesions inhibited further demineralization in vitro, especially in the presence of fluoride. Saliva contamination decreased the ability of the infiltrant to prevent further demineralization, but the presence of fluoride seemed to counteract this effect.

Effect of Gallium on Growth of Streptococcus mutans NCTC 10449 and Dental Tissues

Gallium-doped phosphate-based glasses (Ga-PBG) were assessed for their impact on Streptococcus mutans and dental mineralisation, firstly by disc diffusion assays followed by biofilms grown on nitrocellulose filter membrane (NFM) and constant-depth film fermentor (CDFF). Short-time exposure (10 min) effects of Ga-PBG on S. mutans biofilm were compared with that of 0.2% chlorhexidine. The effects of Ga-PBG on bovine enamel (which was investigated under pH-cycling condition) and dentine were analysed using transverse microradiography (TMR), profilometry and inductively coupled plasma optical-emission spectrometry (ICP-OES). The disc diffusion assays showed inhibition zones of 24.5 ± 0.5 mm for Ga-PBG compared with controls (C-PBG). Ga-PBG showed statistically significant growth inhibition of S. mutans biofilms on NFM (p = 0.001) and CDFF (p < 0.046) compared with hydroxyapatite (HA) and C-PBG. The CDFF assay revealed a maximum of 2.11 log colony-forming unit (CFU) reduction at 48 h, but short-time exposure effects were comparable with that of 0.2% chlorhexidine only on older biofilms (maximum of 0.59 vs. 0.69 log CFU reduction at 120 h). TMR analyses of the enamel revealed non-significant mineral loss (p = 0.37) only in the case of Ga-PBG samples compared with controls including sodium fluoride. ICP-OES analyses indicated transient gallium adsorption into dentine by calcium displacement. The results confirmed that gallium inhibited S. mutans growth and appears to have the potential to protect the enamel surface under conditions representative of the oral environment. Further work is needed to establish whether it has an application in daily oral hygiene procedures to prevent or reduce caries.

Cerium Chloride Reduces Enamel Lesion Initiation and Progression in vitro

Aim: Determination of the potential of cerium chloride to reduce artificial carious mineral loss and lesion depth progression. Methods: A total of 160 enamel samples were prepared from 40 bovine lower central incisors. Crowns were sectioned into four pieces, embedded in acrylic resin, ground flat and allocated to eight groups (S1-S4 and D1-D4; n = 20). Specimens of groups D1-D4 were stored (for 7 days) in a demineralizing buffer solution to induce caries-like lesions. Afterwards, samples were treated for 30 s with one of the following solutions: placebo (S1 and D1), amine fluoride (S2 and D2), cerium chloride (S3 and D3) and a combination of fluoride and cerium chloride (S4 and D4). After another 7 (D1-D4) or 14 (S1-S4) days in demineralizing buffer solution, integrated mineral loss and lesion depth were determined by transversal microradiography and compared by Scheffé's post hoc tests. Results: In groups S1-S4, the highest values for integrated mineral loss and lesion depth were observed for group S1 (placebo), the lowest values for group S4. The results in groups S2-S4 were not significantly different. In groups D1-D4, the highest values for integrated mineral loss and lesion depth were observed for group D1 (placebo), the lowest values in groups D3 and D4. In group D2, integrated mineral loss and lesion depth were significantly lower as compared to D1, but significantly higher compared to groups D3 and D4. Conclusion: Cerium chloride and its combination with fluoride are able to significantly reduce carious mineral loss and the progression of lesion depth.